Department of Transplant Surgery, The Third Xiangya Hospital of Central South University, Changsha 410013, China.
Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, China.
Int J Med Sci. 2019 Jan 24;16(2):343-354. doi: 10.7150/ijms.29393. eCollection 2019.
Ischemic postconditioning (IPO) attenuates hepatic ischemia/reperfusion (I/R) injury. However, little is known about the underlying biological pathophysiology, which could be, at least in part, informed by exploring the transcriptomic changes using next-generation RNA sequencing (RNA-Seq). In this study, 18 mice (C57BL/6) were involved and randomly assigned to three groups: normal (n=6), I/R (n=6, subjected to 70% hepatic I/R), and IR+IPO (n=6, applying IPO to mice with I/R injury). We randomly selected 3 mice per group and extracted their liver tissues for next-generation RNA-Seq. We performed a bioinformatics analysis for two comparisons: normal vs. I/R and I/R vs. IR+IPO. From the analysis, 2416 differentially expressed genes (DEGs) were identified ( < 0.05 and fold change ≥ 1.5). Gene ontology (GO) analysis revealed that these genes were mainly related to cellular metabolic processes, nucleic acids and protein binding processes. The enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways for the DEGs were the mitogen-activated protein kinase (MAPK), IL-17 signalling pathway, regulating pluripotency of stem cells, and insulin resistance pathway. Validation of 12 selected DEGs by qRT-PCR showed that Cyr61, Atf3, Nr4a1, Gdf15, Osgin1, Egr1, Epha2, Dusp1, Dusp6, Gadd45a and Gadd45b were significantly amplified. Finally, a protein-protein interaction (PPI) network constructed to determine interactions of these 11 DEGs. In summary, by exploring gene expression profiling in regard to hepatic I/R and IPO using next-generation RNA-Seq, we suggested a few progression-related genes and pathways, providing some clues for future experimental research.
缺血后处理（IPO）可减轻肝缺血/再灌注（I/R）损伤。然而，对于潜在的生物学病理生理学机制知之甚少，这至少可以部分通过使用下一代 RNA 测序（RNA-Seq）探索转录组变化来提供信息。在这项研究中，涉及了 18 只小鼠（C57BL/6），并将它们随机分为三组：正常组（n=6）、I/R 组（n=6，接受 70%的肝 I/R）和 I/R+IPO 组（n=6，对 I/R 损伤的小鼠应用 IPO）。我们从每组中随机选择 3 只小鼠，提取其肝脏组织进行下一代 RNA-Seq。我们对两个比较进行了生物信息学分析：正常组 vs. I/R 组和 I/R 组 vs. I/R+IPO 组。通过分析，鉴定出 2416 个差异表达基因（DEGs）（ < 0.05，倍数变化≥1.5）。GO 分析表明，这些基因主要与细胞代谢过程、核酸和蛋白质结合过程有关。DEGs 富集的京都基因与基因组百科全书（KEGG）途径为丝裂原激活蛋白激酶（MAPK）、IL-17 信号通路、调节干细胞多能性和胰岛素抵抗途径。通过 qRT-PCR 验证了 12 个选定的 DEGs，结果表明 Cyr61、Atf3、Nr4a1、Gdf15、Osgin1、Egr1、Epha2、Dusp1、Dusp6、Gadd45a 和 Gadd45b 明显扩增。最后，构建了一个蛋白质-蛋白质相互作用（PPI）网络来确定这 11 个 DEGs 的相互作用。总之，通过使用下一代 RNA-Seq 探索肝 I/R 和 IPO 相关的基因表达谱，我们提出了一些与进展相关的基因和途径，为未来的实验研究提供了一些线索。
